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Two Decades on Ice

1050612  Kris Lindsey, CREW Research Associate, and Dr. Dani Ballesteros, CREW Post-doc, removing samples from one of the Frozen Garden’s storage tanks.

Kris Lindsey, CREW Research Associate, and Dr. Dani Ballesteros, CREW Post-doc, removing samples from one of the Frozen Garden’s storage tanks.

This year CREW’s Plant Division is starting work on an exciting project:  Removing samples from the Frozen Garden of CREW’s CryoBioBank, some of which have been frozen in liquid nitrogen at -196oC for up to 25 years.  The official title of the project is:  Evaluating Two Decades of Seeds, Spores, and Tissues in CREW’s CryoBioBank: Cryostorage as a Tool for Ex Situ Conservation in Botanical Gardens, and the work is part of a National Leadership Grant from the Institute of Museum and Library Services (IMLS).  CREW has one of the oldest and most diverse frozen collections of wild plant materials in the world.  The Frozen Garden includes seeds, spores, embryos, gametophytes of mosses and ferns, pollen, and shoot tips from a wide variety of species, many of which are endangered.

The sample box is quickly moved to a small box with liquid nitrogen, to keep all the samples in the box frozen, while some are removed.

The sample box is quickly moved to a small box with liquid nitrogen, to keep all the samples in the box frozen, while some are removed.

Over the course of the next three years we will be removing samples from the collection to test their viability and to look at their genetic integrity.  Close to 1000 samples are targeted for the project—about one-third of the collection.  In addition, some samples that have been frozen in liquid nitrogen have had duplicate samples that were stored under other conditions:  -20oC (which is the temperature most seed banks use), 4oC (refrigerator temperature), and at about 22oC (room temperature).  These will provide valuable comparisons with the samples stored in LN.

Removing the targeted sample from the sample box.  The box with the rest of the samples is then quickly moved back into the storage tank.

Removing the targeted sample from the sample box. The box with the rest of the samples is then quickly moved back into the storage tank.

Working on this project will be a team effort.  Dr. Valerie Pence and Kris Lindsey are being joined by CREW’s new post-doc, Dr. Dani Ballesteros, Megan Philpott, a Ph.D. student and her advisor, Dr. Theresa Culley, from the Department of Biological Sciences at the University of Cincinnati.  Dani will be primarily working on studies of survival, while Megan will be looking at the genetics of the samples.  During the course of the project she will also travel to the USDA-ARS National Center for Genetic Resources Preservation (NCGRP) lab in Ft. Collins, CO, to work with Dr. Christina Walters on DNA degradation studies of the samples.

Megan Philpott, UC Ph.D. student, preparing solutions for DNA isolation.

Megan Philpott, UC Ph.D. student, preparing solutions for DNA isolation.

Removing the samples is a look at the past, reviving plant materials that have been in “suspended animation” for up to two decades.  But, it will also provide valuable information and guidance for the future, as botanical gardens work to preserve the rich plant diversity of the world using the best practices available.  Look for updates as this project unfolds in the coming months.

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